scholarly journals Detection of Zika Virus in Desiccated Mosquitoes by Real-Time Reverse Transcription PCR and Plaque Assay

2017 ◽  
Vol 23 (4) ◽  
pp. 680-681 ◽  
Author(s):  
Kristen L. Burkhalter ◽  
Harry M. Savage
2016 ◽  
Vol 55 (2) ◽  
pp. 535-544 ◽  
Author(s):  
Kirsten St. George ◽  
Inderbir S. Sohi ◽  
Elizabeth M. Dufort ◽  
Amy B. Dean ◽  
Jennifer L. White ◽  
...  

ABSTRACTThe performance and interpretation of laboratory tests for Zika virus (ZKV) continue to be evaluated. Serology is cross-reactive, laborious, and frequently difficult to interpret, and serum was initially solely recommended for molecular diagnosis. ZKV testing was initiated in January 2016 in New York State for symptomatic patients, pregnant women, their infants, and patients with Guillain-Barré syndrome who had traveled to areas with ZKV transmission. Subsequently, eligibility was expanded to pregnant women with sexual partners with similar travel histories. Serum and urine collected within 4 weeks of symptom onset or within 6 weeks of travel were tested with real-time reverse transcription-PCR (RT-PCR) assays targeting the ZKV envelope and NS2B genes. In this review of lessons learned from the first 80 positive cases in NYS, ZKV RNA was detected in urine only in 50 patients, in serum only in 19 patients, and in both samples concurrently in 11 patients, with average viral loads in urine a log higher than those in serum. Among 93 positive samples from the 80 patients, 41 were positive on both gene assays, 52 were positive on the envelope only, and none were positive on the NS2B only. Of the 80 infected patients, test results for 74 (93%) would have defined their infection status as not detected or equivocal if the requirement for positive results from two assay targets (two-target-positive requirement) in the initial federal guidance to public health laboratories was enforced, if urine was not tested, or if the extended eligibility time for molecular testing was not implemented. These changes facilitated more extensive molecular diagnosis of ZKV, reducing reliance on time-consuming and potentially inconclusive serology.


2010 ◽  
Vol 73 (2) ◽  
pp. 400-404 ◽  
Author(s):  
MARYLINE GIRARD ◽  
SOLANGE NGAZOA ◽  
KIRSTEN MATTISON ◽  
JULIE JEAN

The aims of this study were (i) to evaluate the impact of pH and relative humidity on the attachment of norovirus (NoV) to fomites and (ii) to evaluate the effectiveness of different household disinfectants on NoV attached to fomites. Plaque assay and/or real-time reverse transcription PCR assay were used to determine the amount of murine and human NoV attached to stainless steel disks, i.e., the amount removed by sonication in elution buffer but not by surface rinses with water only. An enzymatic pretreatment was used for both human and murine NoV before the real-time reverse transcription PCR assay to avoid detection of RNA associated with inactivated virus. For both murine and human NoV, maximum attachment was obtained after a contact time of 10 min. Attachment of NoV to stainless steel does not appear to be affected by pH, although murine NoV was less attached (<2 log units) at pH 9 and at low relative humidity (25%) than was human NoV (3 log units). Sodium hypochlorite (3%) was the most effective disinfectant, producing a greater than 3-log reduction after 10 min compared with less than a 1-log reduction after treatment with quaternary ammonium compounds and ethoxylated alcohols. Murine NoV was more sensitive than human NoV to disinfectants by approximately 1 to 2 log units. These results will help improve strategies for decontaminating surfaces harboring NoV and thus reduce the incidence of illness caused by these pathogens in the food sector and domestic environments.


2007 ◽  
Vol 73 (7) ◽  
pp. 2338-2340 ◽  
Author(s):  
Yu-Chen Hwang ◽  
Oymon M. Leong ◽  
Wilfred Chen ◽  
Marylynn V. Yates

ABSTRACT Two newly developed protocols for infective virus detection were compared to the plaque assay. An immunomagnetic separation procedure coupled with real-time reverse transcription-PCR of viral nucleic acids was developed to identify intact enteroviral particles, and a reporter cell system responding to viral replication based on fluorescent resonance energy transfer for detection of infectious enteroviruses was tested. Both new procedures detected infective viruses in environmental samples at the same level as the plaque assay.


2007 ◽  
Vol 74 (2) ◽  
pp. 543-546 ◽  
Author(s):  
Leen Baert ◽  
Christiane E. Wobus ◽  
Els Van Coillie ◽  
Larissa B. Thackray ◽  
Johan Debevere ◽  
...  

ABSTRACT The correlation between the detection of murine norovirus 1 RNA by real-time reverse transcription-PCR and the infectivity by plaque assay before and after heat exposure (80°C) was examined. No correlation was found in the current study. Moreover, heat inactivation had a much stronger detrimental effect on virus infectivity than on the integrity of the viral genome.


2021 ◽  
pp. 104868
Author(s):  
Marielle BEDOTTO ◽  
Pierre-Edouard FOURNIER ◽  
Linda HOUHAMDI ◽  
Philippe COLSON ◽  
Didier RAOULT

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